Proteomics

SILAC (Stable Isotopic Labelling of Amino Acids in Cell Culture) (Ong et Mann, 2006) has been proven as a powerful technique for quantitative proteomics in cell culture. The method is robust and provides accurate results (Ong et al., 2002).

The procedure in short:

Cells are grown in culture media that are identical with exception of amino acids. The "light” culture (L) contains all the amino acids in unlabeled form and the “heavy” culture (H) contains one or two amino acids in labeled form. The labeled amino acids are usually lysine labelled with 2H (D4), 13C or 13C15N and/or arginine labeled with 13C or 13C15N. The labeled amino acids increase the molecular weight allowing identification and quantification of the proteins/peptides stemming from culture (L) and (H), respectively.

 

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